EPA Method 552.1 is an ion exchange procedure used for determining haloacetic acids and dalapon in drinking water and drinking water sources.
The purpose of this application note is to present data demonstrating the capability of the Horizon Technology SPE-DEX® 4790 Automated Extraction System to perform the sample extraction required for this method.

In all areas of analytical laboratory testing it is vital to ensure proper quality measures are in place to reduce or eliminate cross contamination between samples, which could result in false positive and/or false negative results. In many cases sample carryover in the LC/MS system is checked early on in the method development process. However, one area that can often be overlooked the sample preparation stage. This involves all aspects including pipetting, sample transfer, extraction protocol, evaporation and mixing steps. This poster examines various stages of the sample preparation process to determine the potential for cross contamination and present approaches to minimize and or eliminate the effect. This is demonstrated via a series of dye experiments combined with analyte testing using LC-MS/MS.

This poster demonstrates the use of a novel protein and phospholipid depletion plate, for the extraction of 25-hydroxy vitamin D from serum. The extraction protocol was ultimately transferred to an SPE automation platform and method performance versus manual processing was compared.
ASMS 2015

The new ISOLUTE® PLD+ Protein and Phospholipid Removal Plate is highlighted in this poster. Protein and phospholipid removal, and analyte recovery are included, along with data illustrating the positive impact of clean (PL and protein free) samples in maintaining analyte signal intensity and low UPLC column back pressure over multiple LCMS runs.
HPLC 2014

This poster evaluates the performance of a novel 96-well filter plate (ISOLUTE PLD+ Protein and Phospholipid Removal Plate) for
the simultaneous removal of proteins and phospholipids from serum samples prior to LC-MS/MS analysis.
ASMS2014

This poster presents the ISOLUTE PLD+ Protein and Phospholipid removal plate, highlighting its ease of use and excellent performance with respect to sample clean up, analyte recovery and elimination of back pressure build up in the UPLC system.
IMSC 2014

Disinfection by-products (DBP) are an ever-present nuisance in the efforts to purify drinking water, wastewater and municipal waters from various sources. An emerging class of DBP compounds with health effects is nitrosamines1-3 which result from chloramination or chlorination if the water is nitrogen-rich.

In postmortem cases, where drugs or pesticides have been used for
their poisonous properties, traditional matrices such as urine and
whole blood may be inappropriate for qualitative and quantitative
analysis. As the site of metabolism for most drugs and toxins, the
liver may provide more insight to cause of death than other bodily
fluids.
This poster describes the use of ISOLUTE SLE+ supported liquid extraction columns to extract a range of drug and pesticide classes form homogenised liver using a simple, streamlined workflow.
SOFT 2016

Matrix components, particularly salts, proteins and
phospholipids, can lead to ion suppression resulting in
inaccurate quantitation and reduced detection limits.
Resin-based mixed-mode cation exchange SPE sorbents
are widely used for the extraction of basic compounds
from biological fluids. The dual retention mechanism
allows a two stage interference wash protocol, which
results in extremely clean extracts.
This poster will investigate the performance of
EVOLUTE™ CX for the extraction of a wide range of basic
drugs from plasma, showing high analyte recovery and
advanced extract cleanliness. The analyte suite was
selected to cover a variety of basic analytes with wide
ranging polarities (LogP values). Extract cleanliness
experiments were performed showing overall ion
suppression, then individual matrix components examined
in terms of protein and phospholipid removal.

This poster evaluates the extraction of a range of drugs of abuse from hydrolyzed and nonhydrolyzed urine using a novel flow through scavenging product, ISOLUTE® HYDRO DME+. Matrix component removal in terms of creatinine and urea, salt residue, pigmentation associated with urobillin content and protein removal will be demonstrated.
SOFT 2018

Methylisothiazolinone (MI) is used in a variety of personal care products, such as sunscreens, lotions, cosmetics. MI is a cytotoxin and as a result there is concern because of sensitization and allergic reactions as well as cell and nerve damage. A percentage of the population is at risk from contact dermatitis when exposed to this compound at sufficient concentrations. This poster describes the use of ISOLUTE SLE+ for extraction of MI from sunscreen.
ASMS 2015

This poster compares the performance of manual processing to a novel automated sample preparation system prior to GC/MS or LC-MS/MS analysis in forensic toxicology applications. Emphasis is placed on the potential for 96-well cross contamination and strategies for its elimination.
TIAFT 2016
SOFT 2016

Matrix components, particularly salts, proteins and phospholipids, can lead to ion suppression resulting in inaccurate quantitation and reduced detection limits.
Resin-based mixed-mode cation exchange SPE sorbents are widely used for the extraction of basic compounds from biological fluids. The dual retention mechanism allows a two stage interference wash protocol, which results in extremely clean extracts. This poster will investigate the performance of EVOLUTE™ CX for the extraction of a wide range of basic
drugs from plasma, showing high analyte recovery and advanced extract cleanliness. The analyte suite was selected to cover a variety of basic analytes with wide ranging polarities (LogP values). Extract cleanliness experiments were performed showing overall ion suppression, then individual matrix components examined
in terms of protein and phospholipid removal.

Amphetamine, methamphetamine and Ecstasy continue to be widely abused in many parts of the world. Urine analysis is the most popular approach to determining drug intake. This poster examines various sample preparation approaches in the analysis of amphetamines prior to gas chromatography-mass spectrometry.
Comparisons were performed between silica-based and polymer-based SPE (ISOLUTE HCX and EVOLUTE EXPRESS CX) as well as supported liquid extraction (SLE, ISOLUTE SLE+).
TIAFT 2019

Traditionally the analysis of aldosterone and angiotensin (for plasma renin activity measurement) are performed separately. However, the relationship of the aldosterone-to-renin-ratio is a very useful tool to help define the cause of secondary hypertension. This poster compares sample preparation options for the simultaneous extraction of Angiotensin I, II and Aldosterone from plasma.
ASMS 2018

Traditionally the analysis of aldosterone and angiotensin (for plasma renin activity measurement) are performed separately. However, the relationship of the aldosterone-to-renin-ratio is a very useful tool to help define the cause of secondary hypertension. This poster compares sample preparation options for the simultaneous extraction of Angiotensin I, II and Aldosterone from plasma.
MSACL EU 2018
MSACL NA 2019

For as long as column chromatography has been practiced by
organic and medicinal chemists there has been a concern that too
much methanol in the solvent system will dissolve the column’s
silica. This concern has come about because chemists often find a
solid white precipitate in collected fractions after purification. This
phenomenon is usually noticed when dichloromethane and
methanol are used as the mobile phase solvents.
In this poster we will show the results of a study that evaluated the
degree of dissolution for both granular and spherical silica in
methanol and dichloromethane.

This poster describes an improved workflow for the analysis of
drugs of abuse from hair. Implementation of bead homogenization along with automated sample preparation allowed for simplified methodology. The direct solvent extraction approach avoids the need for pre-concentration while maintaining desired LOQs with either 200 or
400 μL of hair extract.
SOFT 2018

Sample preparation for hair analysis is often lengthy involving multiple manual labor steps from cutting, washing, homogenization/pulverization, digestion, sample extraction and analysis. This poster aims to demonstrate a streamlined sample
preparation workflow for hair analysis.
The workflow is evaluated using a suite of drugs of abuse, including THC and metabolites.
MSACL NA 2019

This poster presents a simple method for the extraction and subsequent detection of both the traditional hydroxy metabolites and the biologically active dihydroxy metabolites in serum. High analyte recoveries and low ion suppression were demonstrated, allowing limits of quantitation at low pg/mL levels for the di-OH metabolites and < 1 ng/mL levels for the OH metabolites.
MSACL 2014

Most drugs are excreted in urine as glucuronide conjugates. Hydrolysis using a beta-glucuronidase enzyme to convert the metabolites to their “free” form for analysis increases sensitivity. Red abalone (Kura Biotech), abalone (Campbell Scientific), and recombinant (IMCSzyme) beta-glucuronidase enzymes were evaluated to determine which provided the most complete hydrolysis of glucuronide metabolites without effecting the overall recovery of non-conjugated compounds.
EVOLUTE EXPRESS CX 96-well plates were used to extract hydrolysed urine samples, and the impact of th enzymes was compared.
MSACL 2017, Palm Springs
SOFT 2017, Boca Raton

EXPRESS represents a leap forward in high
throughput solid phase extraction. The EVOLUTE
EXPRESS range of 96-well SPE plates combines
powerful EVOLUTE sorbent chemistry with innovative
features that enhance productivity by optimizing
or even eliminating the need for some traditional
steps in the SPE procedure. By truly eliminating the
need for conditioning and equilibration, samples
can be prepared by loading the plate, washing away
interferences and eluting with compatible solvents

EVOLUTE® EXPRESS Solid Phase Extraction products
from Biotage are designed for all your sample
preparation challenges. Whether you are working in the field of bio-analysis during drug development, clinical or forensic
toxicology, or with the diversity of samples in food
safety and environmental applications, the tools and
methods used for sample preparation have a clear
impact on productivity. This user guide contains information about choosing the correct EVOLUTE EXPRESS product for your application and optimizing your methodology for best results.

Sample preparation is generally regarded as a pain point for many labs as it can be laborious, time consuming and relatively expensive. In a high throughput setting performing offline chemistry also leads to increases in sample turnaround time and reporting.
This poster evaluates simplified sample preparation workflows using optimized sample hold-up technology allowing chemistry in-situ. Reduced labour, extraction complexity, processing time, solvent usage and resultant instrument contamination and downtime can offset upfront costs associated with sample preparation.
ASMS 2019

Through innovative patented technology molecularly
imprinted polymers (MIPs) can afford improved clean-up in
your processes. Unwanted contaminants or high value
desirables can be efficiently extracted from processes
streams, resulting in more efficient production and cleaner
products.

Pharmaceutical companies and those who supply them with intermediates, constantly strive to produce API’s and
intermediates of the highest purity. At the same time, many established production facilities face strong competitive
pressures to produce high quality product at lower costs. Drug substance impurities are wide-ranging as described in the
International Conference on Harmonization (ICH) “Impurities in new Drug Substances”1. Of particular concern in recent
years are the ‘genotoxic compounds.

This poster discusses various stages of the sample preparation process to determine and assess the potential for cross contamination and present approaches to minimize and or eliminate the effect.
MSACL 2016

The data given in this Application Note illustrates the fact that the combination of the Biotage(R) Horizon SmartPrep Automated Extractor, DryVap Concentrator, and DryDisk Separation Membranes represent a complete solution to the preparation of samples for the extraction of semi-volatile organics by EPA Method 525.3.

China has set ambitious goals for cleanup and monitoring of drinking water. Methods that are rugged and sensitive are required to support this effort. Chinese Environmental Agency method SL 392-2007 has been developed for this purpose and specifies a C18 cartridge for extraction and GC/MS for the detection step.

This poster describes a comparison of 25-hydroxy Vitamin D Extraction using Supported Liquid Extraction and Phospholipid Depletion Plate Technology using Manual and Automated Sample Preparation prior to LC/MS Analysis. It was presented at MSACL 2015, San Diego

This methodology has been designed to give an effective and efficient supported liquid extraction protocol for the clean-up and concentration of salivary cortisol levels. Cortisol is a steroid hormone that when measured from saliva can be used as an indication of stress. Analyte recoveries achieved using this method ranged from 96-99% with RSDs below 3% (n=7) for all analytes.
Cortisol, Saliva, SLE, Supported Liquid Extraction, ISOLUTE, Clinical, Steroid Hormones, Stress

Cortisol is a steroid hormone that in urine can be used to diagnose hyper or hypo cortisol diseases such as Cushing’s Syndrome. This methodology has been designed to give an effective and efficient supported liquid extraction protocol for the clean-up and concentration of urinary cortisol levels. Analyte recoveries achieved using this method ranged from 99-101% with RSDs below 5% for all analytes.
Cortisol, Steroid hormone, Cushing's Syndrome, Cushing's Disease, Clinical, SLE, Supported Liquid Extraction, Stress,

This application note describes the extraction of 11-nor-9-carboxy-THC from a urine matrix, prior to GC/MS analysis.
Carboxy-THC is the primary metabolite of THC, a key indicator of illicit marijuana usage. In urine, ~80% of the carboxy-THC metabolite is present in the form of its glucuronide metabolite. Therefore, to effectively quantify the THC-COOH, urine is hydrolyzed before extraction. This application note describes optimized extraction of urine samples prepared by either enzymatic or base hydrolysis.

This application note describes the extraction of THC and metabolites from urine using ISOLUTE SLE+ columns or plates. The most prevalent marker in biological samples taken from cannabis abusers is 11-nor-9-carboxy-Δ9-THC. Here we demonstrate a supported liquid extraction procedure for 11-nor-9-carboxy-Δ9-THC.

The use of schedule I drugs for patient pain management therapy warrants constant monitoring of therapeutic levels in patient urine samples. Screening patient urine samples is further complicated by the metabolic process which converts the free drug to the - glucuronide form. The target drugs in patient urine samples can be enzymatically hydrolysed and extracted using Supported Liquid Extraction (ISOLUTE SLE+) which offers an efficient alternative to traditional liquid-liquid extraction (LLE) and solid phase extraction (SPE) techniques typically used for this type of clinical sample preparation.
San Diego, MSACL, 2013

This application note describes a Supported Liquid Extraction (SLE) protocol for the extraction of various drugs of abuse from hydrolyzed urine prior to LC-MS/MS analysis.
drugs of abuse, sle, multiclass

This application note describes a Supported Liquid Extraction (SLE) protocol for the extraction of various drugs of abuse from non- hydrolyzed urine prior to LC-MS/MS analysis.
drugs of abuse, sle, multiclass

This application note describes a SPE-UHPLC-MS/MS procedure for the analysis of 8-oxodG, a biomarker of oxidative stress, from various human biofluids using ISOLUTE ENV+ solid phase extraction columns. Oxidative stress is the imbalance between the production of reactive oxygen species (ROS) and the body’s ability to eliminate them, in favour of the former. ROS have been linked with development of numerous diseases including Parkinson’s and Alzheimer’s and have also been associated with fertility problems. 8-Oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) is one of the mutagenic nucleobase modifications produced in DNA and the 2’deoxyribonucleotide pool, by the reaction of ROS. Measurement of 8-oxodG in biological fluids can provide a useful indicator of oxidative stress.
8-oxodG, sample prep, ENV+, urine, biological fluids, biomarker, oxidative stress, nucleotide,

This application note describes the extraction of a panel of 18 steroid hormones from horse hair using ISOLUTE® SLE+. The simple sample preparation procedure delivers clean extracts and analyte recoveries greater than 80% with RSDs lower than 5% for most analytes. Linearity of greater than 0.999 is achieved for all analytes in the range 0.5-500 pg/mg of hair.

This application note describes the extraction of a panel of 19 steroid hormones from human serum using EVOLUTE® EXPRESS ABN solid phase extraction plates prior to LC/MS analysis. The simple sample preparation procedure delivers clean extracts and analyte recoveries greater than 80% with RSDs lower than 10% for the majority of analytes. Linearity of >0.99 is achieved for all analytes in the range 5–5000 pg/mL.
The application note includes details on automation of the extraction method using Biotage® Extrahera.

This application note describes the extraction of 47 drugs of abuse and metabolites from oral fluid matrix collected using the Intercept Oral Fluid Drug Test Kit (Orasure Technologies), prior to UPLC-MS/MS analysis. The sample preparation is optimised to minimise matrix effects due to the buffers used in the collection device. Estimated LOQs range from 0.1-1 ng/mL for the various analytes.

Drinking water is a significant source of environmental exposure, especially for small children. Countries around the world have put regulations in place to monitor drinking water quality for a wide range of hazardous compounds. Methods such as SL 392-2007 in China, the EN methods in Europe and US methods such as method 525.2 cover a large suite of analytes of concern. They can be effectively extracted using solid phase extraction (SPE) disks and using GC/MS for detection.

US EPA method 525.2 may use solid phase extraction (SPE) to extract the analytes of interest from water samples. It includes a variety of quality control measures to ensure the method is under control throughout the analysis. The Biotage® Horizon 5000 was used in this study to extract the EPA Method 525.2 analytes from six prepared water samples.
The Atlantic high-capacity C18 disks provided excellent recovery of the large suite of compounds extracted in water. The Biotage® Horizon 5000 system provided uniform performance and a hands-off approach to the extraction step. The reproducibility of the six runs was excellent and the average of the standard deviation values was 2.57%.

This application note describes the use of the ISOLUTE® PLD+ Protein and Phospholipid Removal Plate for clean-up of a range of acidic, basic and neutral drugs from plasma. ISOLUTE® PLD+ Protein and Phospholipid Removal Plates are suitable for clean-up of a range of analytes with widely differing functionality and polarity characteristics from plasma, giving high recoveries, good reproducibility and excellent extract cleanliness.

This methodology has been designed to give an effective and efficient supported liquid extraction protocol for the clean-up and concentration of a range of forensically significant amphetamines followed by derivatization to optimize for GC-MS analysis. Analyte recoveries achieved using this method ranged from 99-104% with RSDs below 10% for all analytes.
Amphetamines, SLE, Supported Liquid Extraction, SLE+, ISOLUTE, Urine, GC, GC-MS,

Immunosuppressants inhibit or prevent the activity of the immune system used to prevent rejection of transplanted organs and treat autoimmune diseases. Due to the risks of immunodeficiency, those under treatment must undergo constant therapeutic drug monitoring (TDM) requiring reliable and robust analytical techniques for quantification of these drugs. Current methodologies use immunoassay techniques to measure immunosuppressant levels in patients which are expensive, time consuming and susceptible to issues with cross reactivity. The sample prep method in this application note offers an alternative approach to immunosuppressant analysis with LC-MS/MS giving more reliable and robust recoveries with no cross-reactivity and cost saving opportunities. Analyte recoveries range from 60-97% with RSDs all below 10%.
Immunosuppressant, Tacrolimus, Sacrolimus, Everolimus, Cyclosporin A, Clinical, SLE+, SLE, Supported Liquid Extraction, immune, Clinical, Whole Blood, immunoassay,

This application note demonstrates an effective and efficient suported liquid extraction protocol for the clean up and concentration of a range of forensically significant opiates and their metabolites
opiates, sle, urine, GC-MS, forensic, drugs of abuse, DOA, UCT, agilent

This application note describes a solid phase extraction protocol for the extraction of three catecholamine metabolites (vanillylmandelic acid, homovanillic acid and 5-hydroxyindoleacetic acid) from plasma prior to LC-MS/MS detection. The method described in this application achieves high reproducible recoveries and excellent linearity (>0.99) in the range 10-200 ng/mL.

This application note describes a supported liquid extraction protocol for the extraction of three acidic catecholamine metabolites (vanillylmandelic acid, homovanillic acid and 5-hydroxyindoleacetic acid) from plasma prior to LC-MS/MS detection. The method described in this application note achieves high reproducible recoveries (>80%, RSD < 5%) for these analytes in plasma with linearity > 0.995 in the range
2–200 ng/mL.

This application note demonstrates an effective and efficient suported liquid extraction protocol for the clean up and concentration of a range of forensically significant opiates and their metabolites
ISOLUTE, unknown screening,

The method addresses the extraction of all three classes of drugs of abuse from urine using non-polar and cation exchange retention mechanisms on the ISOLUTE HCX mixed mode column. This protocol was optimized to eliminate the use of dichloromethane in the elution of the acidic / neutral drug fraction.

This poster demonstrates a novel, sensitive, cost effective and rugged method for the analysis of acrylamide in coffee and potato chips (crisps) using supported liquid extraction prior to LC-MS/MS
analysis. It demonstrates extraction from coffee down
to a concentration of 1 ng/mL and in potato chips to a level
of 10 ng/g (ppb).
ASMS 2014

This application note describes a Supported Liquid Extraction (SLE) protocol for the extraction of acrylamide from coffee using ISOLUTE® SLE+ columns with LC-MS/MS detection.
The method described in this application note achieves high recoveries of acrylamide in coffee. The method is sensitive enough to measure levels as low as 1 ng/mL in coffee (solution), 25 ppb in ground coffee (solid) or 125 ppb in instant coffee (solid, traditional or decaffeinated) and gives good selectivity from what is a challenging matrix.

This application note describes an SPE protocol appropriate for LC-MS/MS analysis of aflatoxin M1 found in infant formula.
The method described in this application note achieves high recoveries of aflatoxin M1 from infant formula with %RSDs and LOQs that all meet the requirements set in European regulations for its measurement.

This application note describes a solid phase extraction(SPE) protocol for the extraction of a range of mycotoxins from dried chili (pimiento) using ISOLUTE® Myco with LC-MS/MS analysis.
The method achieves high recoveries of aflatoxins and ochratoxin from dried chili (pimiento) with %RSDs and LOQs that meet the requirements set in European Union regulations for measurement of these analytes.

This application note describes the extraction of a range of amphetamine-type compounds from whole blood, prior to GC/MS analysis. A protocol that allows the simultaneous extraction of various other drugs of abuse classes: barbiturates, benzodiazepines, cocaine and opiates, is also evaluated.
ISOLUTE® SLE+ columns with 1 mL sample capacity are used to extract whole blood samples following a straightforward sample dilution. No protein precipitation or other pre-treatment is required prior to sample loading. The sample preparation procedure delivers clean extracts, good recoveries and RSD values and LLOQs from 10 ng/mL (analyte dependant).

This application note describes the extraction of a range of amphetamines and metabolites from urine using supported liquid extraction and subsequent analysis by GC/MS. Prior to extraction, a simple oxidation step is performed to eliminate sympathomimetic compounds such as ephedrine and pseudoephedrine so they do not interfere with quantitation of methamphetamine.

This method describes the use of ISOLUTE SLE+ in 96 well plate format for the extraction of a range of amphetamines. This procedure is effective with sample volumes of 100 μL for analyte recoveries > 90%. For larger volume (500 μL) samples, see AN746.
amphetamines, AN742, ISOLUTE, SLE+, DOA, DRUGS OF ABUSE, FORENSIC

The following method(s) have been developed for the class-selective extraction of Amphetamine, Methamphetamine, Phentermine, MDA, MDMA, MDEA from human urine. The method is highly reproducible and offers an average recovery greater than 80%.
MIP, AFFINILUTE, SupelMIP, BIOTAGE, Amphetamines,

This method describes the use of ISOLUTE SLE+ in column format for the extraction of a range of amphetamines. This method is effective for sample volumes of 500μL with analyte recoveries > 95%.
Amphetamines, ISOLUTE, DOA, DRUGS OF ABUSE, SLE+, DRUG SCREEN

This procedure is recommended for the extraction of extensive panel of significant therapeutic and illicit drugs of abuse from whole blood and urine using mixed-mode cation exchange SPE columns. ISOLUTE HCX is a silica based mixed-mode sorbent that combines non-polar and strong cation exchange retention mechanisms for extraction of basic drugs from biological fluids.
Ion exchange, Drugs of Abuse, DOA, Clinical,

Ethylestrenol and stanozolol are anabolic steroids which can be used to increase muscle mass and enhance performance. These drugs have been linked to instances of doping in race horses.
This application note describes a Supported Liquid Extraction (SLE) protocol for the extraction of ethylestrenol and stanozolol from horse urine prior to LC-MS/MS analysis.
The method achieves high reproducible analyte recoveries from both gelding and filly urine. Protocols for 48-well, 96-well plate and column formats are described.

Antiepileptic drugs are prescribed to suppress seizures in epilepsy patients. A variety of different types of AEDs have been synthesized to pharmacologically address different types of epilepsy. The ability to therapeutically monitor these drugs in patients is necessary for maintaining optimal medical care and managing any adverse effects of thedrug. A fast and clean extraction method is needed that works in a variety of biological
matrices and affords a high throughput workflow. In this poster, ISOLUTE SLE+ is demonstrated as an effective way to extract AEDs from serum, oral fluid and urine with good efficiency and recovery. The method was developed on a 96 well plate format to facilitate a high throughput workflow model.
MSACL 2014

In this poster, ISOLUTE SLE+ is demonstrated as an effective way to extract antiepileptic drugs from serum, oral fluid and urine with good efficiency and recovery. The method was developed on a 96-well plate format to facilitate a high throughput workflow model.
ASMS 2014

The ability to therapeutically monitor antiepileptic drugs in patients is necessary for maintaining optimal medical care and managing any adverse effects of the drug. A fast and clean extraction method is needed that works in a variety of biological
matrices and affords a high throughput workflow. Here ISOLUTE SLE+ is demonstrated as an effective way to extract AEDs from serum and urine with good efficiency. The method was developed using a 96-well plate format to facilitate a high throughput workflow model.

About Biotage

Biotage is a global Life Science company that develops innovative and effective solutions for separation within organic and analytical chemistry, as well as for industrial applications. Biotage is listed on NASDAQ Stockholm